Diversity of luciferase sequences and bioluminescence production in Baltic Sea Alexandrium ostenfeldii

The toxic dinoflagellate Alexandrium ostenfeldii is the only bioluminescent bloom-forming phytoplankton in coastal waters of the Baltic Sea. We analysed partial luciferase gene (lcf) sequences and bioluminescence production in Baltic A. ostenfeldii bloom populations to assess the distribution and consistency of the trait in the Baltic Sea, and to evaluate applications for early detection of toxic blooms. Lcf was consistently present in 61 Baltic Sea A. ostenfeldii strains isolated from six separate bloom sites. All Baltic Sea strains except one produced bioluminescence. In contrast, the presence of lcf and the ability to produce bioluminescence did vary among strains from other parts of Europe. In phylogenetic analyses, lcf sequences of Baltic Sea strains clustered separately from North Sea strains, but variation between Baltic Sea strains was not sufficient to distinguish between bloom populations. Clustering of the lcf marker was similar to internal transcribed spacer (ITS) sequences with differences being minor and limited to the lowest hierarchical clusters, indicating a similar rate of evolution of the two genes. In relation to monitoring, the consistent presence of lcf and close coupling of lcf with bioluminescence suggests that bioluminescence can be used to reliably monitor toxic bloom-forming A. ostenfeldii in the Baltic Sea.Peer reviewe

Determination of optical markers of cyanobacterial physiology from fluorescence kinetics

Compared to other methods to monitor and detect cyanobacteria in phytoplankton populations, fluorometry gives rapid, robust and reproducible results and can be used in situ. Fluorometers capable of providing biomass estimates and physiological information are not commonly optimized to target cyanobacteria. This study provides a detailed overview of the fluorescence kinetics of algal and cyanobacterial cultures to determine optimal optical configurations to target fluorescence mechanisms that are either common to all phytoplankton or diagnostic to cyanobacteria. We confirm that fluorescence excitation channels targeting both phycocyanin and chlorophyll a associated to the Photosystem II are required to induce the fluorescence responses of cyanobacteria. In addition, emission channels centered at 660, 685 and 730 nm allow better differentiation of the fluorescence response between algal and cyanobacterial cultures. Blue-green actinic light does not yield a robust fluorescence response in the cyanobacterial cultures and broadband actinic light should be preferred to assess the relation between ambient light and photosynthesis. Significant variability was observed in the fluorescence response from cyanobacteria to the intensity and duration of actinic light exposure, which needs to be taken into consideration in field measurements

Optimising Multispectral Active Fluorescence to Distinguish the Photosynthetic Variability of Cyanobacteria and Algae

This study assesses the ability of a new active fluorometer, the LabSTAF, to diagnostically assess the physiology of freshwater cyanobacteria in a reservoir exhibiting annual blooms. Specifically, we analyse the correlation of relative cyanobacteria abundance with photosynthetic parameters derived from fluorescence light curves (FLCs) obtained using several combinations of excitation wavebands, photosystem II (PSII) excitation spectra and the emission ratio of 730 over 685 nm (Fo(730/685)) using excitation protocols with varying degrees of sensitivity to cyanobacteria and algae. FLCs using blue excitation (B) and green–orange–red (GOR) excitation wavebands capture physiology parameters of algae and cyanobacteria, respectively. The green–orange (GO) protocol, expected to have the best diagnostic properties for cyanobacteria, did not guarantee PSII saturation. PSII excitation spectra showed distinct response from cyanobacteria and algae, depending on spectral optimisation of the light dose. Fo(730/685), obtained using a combination of GOR excitation wavebands, Fo(GOR, 730/685), showed a significant correlation with the relative abundance of cyanobacteria (linear regression, p-value < 0.01, adjusted R2 = 0.42). We recommend using, in parallel, Fo(GOR, 730/685), PSII excitation spectra (appropriately optimised for cyanobacteria versus algae), and physiological parameters derived from the FLCs obtained with GOR and B protocols to assess the physiology of cyanobacteria and to ultimately predict their growth. Higher intensity LEDs (G and O) should be considered to reach PSII saturation to further increase diagnostic sensitivity to the cyanobacteria component of the community

The Role of Citizen Science in Promoting Ocean and Water Literacy in School Communities: The ProBleu Methodology

Human activities continue to degrade oceanic, coastal and inland waters. The generational change in the role of society in actively looking after the health of water resources can be achieved through the expansion of ocean and water literacy in schools. The Network of European Blue Schools established under the EU4Ocean Coalition for Ocean Literacy has improved ocean and water literacy; however, this Network needs to grow and be supported. Here, we present ProBleu, a recently funded EU project that will expand and support the Network, partly through the use of citizen science. The core of the proposed methodology is facilitating school activities related to ocean and water literacy through funding calls to sustain and enrich current school activities, and kick-start and support new activities. The outcomes of the project are anticipated to have widespread and long-term impacts across society, and oceanic, coastal and inland water environments

A data driven approach to flag land affected signals in satellite derived water quality from small lakes

The land-affected signal in remotely sensed radiance from nearshore waters is a common problem for remote sensing, introducing uncertainty in atmospheric correction and subsequent water quality constituent concentration estimates. This study proposes a new method for identifying effects of land on satellite remote sensing of water quality. The new optical water types (OWT) containing the land-affected signal were derived from POLYMER-corrected imagery of the Medium Resolution Imaging Spectrometer in reduced resolution (MERIS RR) and Sentinel-3 Ocean and Land Colour Instrument (OLCI). These were then applied, as part of a larger set of existing OWTs corresponding to the variability observed in natural waters, to satellite images. The ability to identify pixels containing both water and land, and those contaminated with radiance from adjacent land, was evaluated. Our test sites include dark lakes of varying size in Sweden (Lakes Rusken, Bolmen, Ringsjön, and Ivösjön) where the classification showed high sensitivity to land near the lake shore. The land-affected signal is shown to lead to underestimations of chlorophyll-a concentration and Forel-Ule colour indices, and overestimations of turbidity in these lakes, which can be corrected after masking out the land-affected pixels. The land-affected signal is strongest in summer, both NDVI and sun zenith angle covaried with the seasonal variation of land-affected signal. Further, the results confirmed that satellite images with coarser spatial resolution are more prone to land-affected signal compared to images with finer spatial resolution, for small inland water bodies. We propose a data-driven approach for water quality processing with ‘land-affected water types’ as an effective way to improve the lake optical water quality monitoring from water colour sensors

Meta-classification of remote sensing reflectance to estimate trophic status of inland and nearshore waters

Common aquatic remote sensing algorithms estimate the trophic state (TS) of inland and nearshore waters through the inversion of remote sensing reflectance (Rrs ()) into chlorophyll-a (chla) concentration. In this study we present a novel method that directly inverts Rrs () into TS without prior chla retrieval. To successfully cope with the optical diversity of inland and nearshore waters the proposed method stacks supervised classification algorithms and combines them through meta-learning. We demonstrate the developed methodology using the waveband configuration of the Sentinel-3 Ocean and Land Colour Instrument on 49 globally distributed inland and nearshore waters (567 observations). To assess the performance of the developed approach, we compare the results with TS derived through optical water type (OWT) switching of chla retrieval algorithms. Meta-classification of TS was on average 6.75% more accurate than TS derived via OWT switching of chla algorithms. The presented method achieved 90% classification accuracies for eutrophic and hypereutrophic waters and was 12% more accurate for oligotrophic waters than derived through OWT chla retrieval. However, mesotrophic waters were estimated with lower accuracy from both our developed method and through OWT chla retrieval (52.17% and 46.34%, respectively), highlighting the need for improved base algorithms for low - moderate biomass waters. Misclassified observations were characterised by highly absorbing and/or scattering optical properties for which we propose adaptations to our classification strategy

A data driven approach to flag land affected signals in satellite derived water quality from small lakes

The land-affected signal in remotely sensed radiance from nearshore waters is a common problem for remote sensing, introducing uncertainty in atmospheric correction and subsequent water quality constituent concentration estimates. This study proposes a new method for identifying effects of land on satellite remote sensing of water quality. The new optical water types (OWT) containing the land-affected signal were derived from POLYMER-corrected imagery of the Medium Resolution Imaging Spectrometer in reduced resolution (MERIS RR) and Sentinel-3 Ocean and Land Colour Instrument (OLCI). These were then applied, as part of a larger set of existing OWTs corresponding to the variability observed in natural waters, to satellite images. The ability to identify pixels containing both water and land, and those contaminated with radiance from adjacent land, was evaluated. Our test sites include dark lakes of varying size in Sweden (Lakes Rusken, Bolmen, Ringsjön, and Ivösjön) where the classification showed high sensitivity to land near the lake shore. The land-affected signal is shown to lead to underestimations of chlorophyll-a concentration and Forel-Ule colour indices, and overestimations of turbidity in these lakes, which can be corrected after masking out the land-affected pixels. The land-affected signal is strongest in summer, both NDVI and sun zenith angle covaried with the seasonal variation of land-affected signal. Further, the results confirmed that satellite images with coarser spatial resolution are more prone to land-affected signal compared to images with finer spatial resolution, for small inland water bodies. We propose a data-driven approach for water quality processing with ‘land-affected water types’ as an effective way to improve the lake optical water quality monitoring from water colour sensors

A Bayesian approach for remote sensing of chlorophyll-a and associated retrieval uncertainty in oligotrophic and mesotrophic lakes

Satellite remote sensing of chlorophyll-a concentration (chla) in oligotrophic and mesotrophic lakes faces uncertainties from sources such as atmospheric correction, complex inherent optical property compositions, and imperfect algorithmic retrieval. To improve chla estimation in oligo- and mesotrophic lakes, we developed Bayesian probabilistic neural networks (BNNs) for the Sentinel-3 Ocean and Land Cover Instrument (OLCI) and Sentinel-2 MultiSpectral Imager (MSI). The BNNs were built using an in situ dataset of oligo- and mesotrophic water bodies (1755 observations from 178 systems; median chla: 5.11 mg m−3, standard deviation: 10.76 mg m−3) and provide a per-pixel uncertainty percentage associated with retrieved chla. Shifts of oligo- and mesotrophic systems into the eutrophic regime, characterised by higher biomass levels, are widespread. To account for phytoplankton biomass fluctuation, a set of eutrophic lakes (167 observations from 31 systems) were included in this study (maximum chla 68 mg m−3). The BNNs were evaluated through five assessments including single day and time series match-ups with OLCI and MSI. OLCI BNN accuracy gains of >25% and MSI BNN accuracy gains of >15% were achieved in the assessments when compared to chla reference algorithms for oligotrophic waters (chla ≤ 8 mg m−3). In comparison to the reference algorithms, the accuracy gains of the BNNs decreased as chla and trophic levels increased. To measure the quality of the provided BNN uncertainty estimate, we calculated the prediction interval coverage probability (PICP), Sharpness and mean absolute calibration difference (MACD) metrics. The associated BNN chla uncertainty estimate included the reference in situ chla values for most observations (PICP ≥ 75%) across the different performance assessments. Further analysis showed that the BNN chla uncertainty estimate was not constantly well-calibrated across different evaluation strategies (Sharpness 1.7–6, MACD 0.04–0.25). BNN uncertainties were used to test two chla improvement strategies: 1) identifying and filtering uncertain chla estimates using scene-specific thresholds, and 2) selecting the most accurate prior atmospheric correction algorithm per individual satellite observation to retain chla with the lowest BNN uncertainty. Both strategies increased the quality of the chla result and demonstrated the significance of uncertainty estimation. This study serves as research on Bayesian machine learning for the estimation and visualisation of chla and associated retrieval uncertainty to develop harmonised products across OLCI and MSI for small and large oligo- and mesotrophic lakes

Impact of Spectral Resolution on Quantifying Cyanobacteria in Lakes and Reservoirs: A Machine-Learning Assessment

Cyanobacterial harmful algal blooms are an increasing threat to coastal and inland waters. These blooms can be detected using optical radiometers due to the presence of phycocyanin (PC) pigments. The spectral resolution of best-available multispectral sensors limits their ability to diagnostically detect PC in the presence of other photosynthetic pigments. To assess the role of spectral resolution in the determination of PC, a large ( N=905 ) database of colocated in situ radiometric spectra and PC are employed. We first examine the performance of selected widely used machine-learning (ML) models against that of benchmark algorithms for hyperspectral remote sensing reflectance ( Rrs ) spectra resampled to the spectral configuration of the Hyperspectral Imager for the Coastal Ocean (HICO) with a full-width at half-maximum (FWHM) of < 6 nm. Results show that the multilayer perceptron (MLP) neural network applied to HICO spectral configurations (median errors < 65%) outperforms other ML models. This model is subsequently applied to Rrs spectra resampled to the band configuration of existing satellite instruments and of the one proposed for the next Landsat sensor. These results confirm that employing MLP models to estimate PC from hyperspectral data delivers tangible improvements compared with retrievals from multispectral data and benchmark algorithms (with median errors between ∼73 % and 126%) and shows promise for developing a globally applicable cyanobacteria measurement approach

Advancing cyanobacteria biomass estimation from hyperspectral observations: Demonstrations with HICO and PRISMA imagery

Retrieval of the phycocyanin concentration (PC), a characteristic pigment of, and proxy for, cyanobacteria biomass, from hyperspectral satellite remote sensing measurements is challenging due to uncertainties in the remote sensing reflectance (∆Rrs) resulting from atmospheric correction and instrument radiometric noise. Although several individual algorithms have been proven to capture local variations in cyanobacteria biomass in specific regions, their performance has not been assessed on hyperspectral images from satellite sensors. Our work leverages a machine-learning model, Mixture Density Networks (MDNs), trained on a large (N = 939) dataset of collocated in situ chlorophyll-a concentrations (Chla), PCs, and remote sensing reflectance (Rrs) measurements to estimate PC from all relevant spectral bands. The performance of the developed model is demonstrated via PC maps produced from select images of the Hyperspectral Imager for the Coastal Ocean (HICO) and Italian Space Agency's PRecursore IperSpettrale della Missione Applicativa (PRISMA) using a matchup dataset. As input to the MDN, we incorporate a combination of widely used band ratios (BRs) and line heights (LHs) taken from existing multispectral algorithms, that have been proven for both Chla and PC estimation, as well as novel BRs and LHs to increase the overall cyanobacteria biomass estimation accuracy and reduce the sensitivity to ∆Rrs. When trained on a random half of the dataset, the MDN achieves uncertainties of 44.3%, which is less than half of the uncertainties of all viable optimized multispectral PC algorithms. The MDN is notably better than multispectral algorithms at preventing overestimation on low (10 mg m−3). According to our extensive assessments, the developed model is anticipated to enable practical PC products from PRISMA and HICO, therefore the model is promising for planned hyperspectral missions, such as the Plankton Aerosol and Cloud Ecosystem (PACE). This advancement will enhance the complementary roles of hyperspectral radiometry from satellite and low-altitude platforms for quantifying and monitoring cyanobacteria harmful algal blooms at both large and local spatial scales